PMID- 28407382 OWN - NLM STAT- MEDLINE DCOM- 20170921 LR - 20170921 IS - 1365-2958 (Electronic) IS - 0950-382X (Linking) VI - 104 IP - 6 DP - 2017 Jun TI - Carbohydrate uptake in Advenella mimigardefordensis strain DPN7(T) is mediated by periplasmic sugar oxidation and a TRAP-transport system. PG - 916-930 LID - 10.1111/mmi.13692 [doi] AB - In this study, we investigated an SBP (DctP(Am) ) of a tripartite ATP-independent periplasmic transport system (TRAP) in Advenella mimigardefordensis strain DPN7(T) . Deletion of dctP(Am) as well as of the two transmembrane compounds of the tripartite transporter, dctQ and dctM, impaired growth of A. mimigardefordensis strain DPN7(T) , if cultivated on mineral salt medium supplemented with d-glucose, d-galactose, l-arabinose, d-fucose, d-xylose or d-gluconic acid, respectively. The wild type phenotype was restored during complementation studies of A. mimigardefordensis DeltadctP(Am) using the broad host vector pBBR1MCS-5::dctP(Am) . Furthermore, an uptake assay with radiolabeled [(14) C(U)]-d-glucose clearly showed that the deletion of dctP(Am) , dctQ and dctM, respectively, disabled the uptake of this aldoses in cells of either mutant strain. Determination of K(D) performing thermal shift assays showed a shift in the melting temperature of DctP(Am) in the presence of d-gluconic acid (K(D) 11.76 +/- 1.3 microM) and the corresponding aldonic acids to the above-mentioned carbohydrates d-galactonate (K(D) 10.72 +/- 1.4 microM), d-fuconic acid (K(D) 13.50 +/- 1.6 microM) and d-xylonic acid (K(D) 8.44 +/- 1.0 microM). The sugar (glucose) dehydrogenase activity (E.C.1.1.5.2) in the membrane fraction was shown for all relevant sugars, proving oxidation of the molecules in the periplasm, prior to transport. CI - (c) 2017 John Wiley & Sons Ltd. FAU - Meinert, Christina AU - Meinert C AUID- ORCID: 0000-0002-1911-3385 AD - Institut fur Molekulare Mikrobiologie und Biotechnologie, Westfalische Wilhelms-Universitat, Munster, D-48149, Germany. FAU - Senger, Jana AU - Senger J AD - Institut fur Molekulare Mikrobiologie und Biotechnologie, Westfalische Wilhelms-Universitat, Munster, D-48149, Germany. FAU - Witthohn, Marco AU - Witthohn M AD - Institut fur Molekulare Mikrobiologie und Biotechnologie, Westfalische Wilhelms-Universitat, Munster, D-48149, Germany. FAU - Wubbeler, Jan Hendrik AU - Wubbeler JH AD - Institut fur Molekulare Mikrobiologie und Biotechnologie, Westfalische Wilhelms-Universitat, Munster, D-48149, Germany. FAU - Steinbuchel, Alexander AU - Steinbuchel A AD - Institut fur Molekulare Mikrobiologie und Biotechnologie, Westfalische Wilhelms-Universitat, Munster, D-48149, Germany. AD - Environmental Science Department, King Abdulaziz University, Jeddah, Saudi Arabia. LA - eng PT - Journal Article DEP - 20170509 PL - England TA - Mol Microbiol JT - Molecular microbiology JID - 8712028 RN - 0 (Bacterial Proteins) RN - 0 (Carbohydrates) RN - 0 (Gluconates) RN - 0 (Membrane Transport Proteins) RN - 0 (Propionates) RN - 0 (Sugar Acids) RN - 0 (Symporters) RN - A1TA934AKO (Xylose) RN - IY9XDZ35W2 (Glucose) RN - R4R8J0Q44B (gluconic acid) RN - X2RN3Q8DNE (Galactose) SB - IM MH - Alcaligenaceae/genetics/*metabolism MH - Bacterial Proteins/genetics MH - Carbohydrates MH - Galactose/metabolism MH - Gluconates/metabolism MH - Glucose/metabolism MH - Membrane Transport Proteins/genetics/*metabolism MH - Periplasm/physiology MH - Propionates/metabolism MH - Sequence Analysis, DNA MH - Sugar Acids/*metabolism MH - Symporters/metabolism MH - Xylose/metabolism EDAT- 2017/04/14 06:00 MHDA- 2017/09/22 06:00 CRDT- 2017/04/14 06:00 PHST- 2017/04/10 00:00 [accepted] PHST- 2017/04/14 06:00 [pubmed] PHST- 2017/09/22 06:00 [medline] PHST- 2017/04/14 06:00 [entrez] AID - 10.1111/mmi.13692 [doi] PST - ppublish SO - Mol Microbiol. 2017 Jun;104(6):916-930. doi: 10.1111/mmi.13692. Epub 2017 May 9.